摘要
目的研究澳门红树林植物内生放线菌多样性及新颖性,为放线菌新物种及新抗生素发现奠定基础。方法 12份采集自澳门红树林的植物样品,消毒粉碎后,撒在10种分离培养基上分离内生放线菌,采用ISP2培养基进行菌株纯化;通过PCR扩增和测序获得16S r RNA基因序列,并提交到数据库中进行搜索比对,开展内生放线菌多样性和新颖性分析;放线菌潜在新物种经液体发酵、离心后,上清液经乙酸乙酯萃取,菌丝经丙酮浸泡,分别获得发酵液水相样品、发酵液酯相样品和菌丝丙酮浸提液3类样品;样品通过纸片扩散法进行抗菌活性初筛。结果从12份植物样品中得到192株内生放线菌,分布于8个目17个科30个属,其中链霉菌为优势菌属;16S r RNA基因序列相似性低于98.6%的菌株共有22株,其中菌株4Q3S-3和2Q3S-4-2分别为弗莱德门菌属(Friedmanniella)和中村菌属(Nakamurella)的新种,放线菌潜在新物种的鉴定工作正在进行;2株放线菌新种及6株潜在放线菌新种的抗菌活性筛选结果表明,5株具有抗菌活性。结论澳门红树林植物内生放线菌资源丰富多样,新颖性高,具有从中发现放线菌新物种和新抗生素的潜力,值得深入研究。
Objective To study the diversity and novelty of endophytic actinobacteria of the mangrove plants in Macao, and lay the foundation of finding new actinobacterial taxa and antibiotics. Methods After surface sterilization, each sample of twelve mangrove plant samples collected in Macao was grinded into powders, and then spread on ten different kinds of isolation media for strain isolation. Colonies were picked up and streaked on the ISP2 agar to obtain the pure isolates. To analyze the diversity and novelty of endophytic actinobacteria, sequences of 16S rRNA gene obtained by PCR amplification were submitted to gene database for BLAST search. To test antimicrobial activity by disk diffusion method, the fermentation broths and mycelia of the potential new actinobacterial strains were extracted by ethyl acetate and acetone, respectively. Three different samples were obtained, including the organic layer, water layer and acetone extraction. Results A total of 192 endophytic actinobacterial strains were obtained, they distributed in 30 genera affiliated to 17 families of 8 orders. The dominant genus was Streptomyces. The 16S rRNA gene sequence similarities between 22 strains and the related species were all less than 98.6%. Among them, both strain 4Q3S-3 and 2Q3S-4-2 have been already proved to be novel species of the genera Friedmanniella and Nakamurella, respectively. Clarifying the taxonomic positions of the potential new actinobacterial taxa is still in progress. Among two new actinobacterial strains and six potential new actinobacterial strains, five strains exhibited antimicrobial activity. Conclusion The study demonstrated that endophytic actinobacteria from mangrove plants of Macao were high diverse and novel, and thus deserved to be further explored to discover new actinobacterial taxa and antibiotics.
作者
李飞娜
潘臻
庹利
刘少伟
左璇
雷晨
李铭源
孙承航
Li Fei-na Pan Zhen Tuo Li Liu Shao-wei Zuo Xuan Lei Chen Li Ming-yuan Sun Cheng-hang(Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Beijing 100050 Zunyi Medical University, Zunyi 563003 China Pharmaceutical University, Nanjing 211198 University of Macao, Macao 999078)
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2017年第4期284-293,共10页
Chinese Journal of Antibiotics
基金
国家自然科学基金(No.81373308和No.81321004)
澳门大学研究基金(No.MYRG2016-00056-FST)
关键词
红树林
内生放线菌
多样性
新颖性
抗生素
Mangrove
Endophytic actinobacteria
Diversity
Novelty
Antibiotics
